Methods of controlling the breeding behavior of butterflies

ABSTRACT

Methods of controlling the feeding and/or breeding behavior of a target insect are disclosed in which a host plant is provided a substantial distance from a plant of interest, chemical attractant that induces the target insect to lay eggs on the host plant is applied to the host plant, application of the chemical attractant to the host plant is repeated as required to induce residual target-insect populations to lay eggs on the host plant, and wherein the behavior is controlled when a substantial number of offspring belonging to subsequent generations themselves mature and display a preference for laying eggs on the same type of host plant on which they were reared without further application of the chemical attractant is provided. Also disclosed are methods of deterring target insect feeding and breeding on plants of interest by applying toxic plant extract to the plants of interest.

CROSS-REFERENCE TO RELATED APPLICATIONS

This application is a continuation of U.S. patent application Ser. No.13/576,080, filed Oct. 22, 2012, entitled “Methods of Controlling theBreeding Behavior of Butterflies”; which is a National Phase applicationof International Application No. PCT/IB11/00440, filed Jan. 28, 2011,entitled “Methods of Controlling the Breeding Behavior of Butterflies”;which claims the benefit of priority to U.S. Provisional Application No.61/299,125, filed Jan. 28, 2010, entitled “Methods of Controlling theBreeding Behavior of Butterflies”, the content of which are herebyincorporated by reference, in their entirety.

BACKGROUND

The Citrus Swallowtail butterfly, Heraclides andraemon Hubner, was firstobserved in Jamaica in 1945. Since its arrival, it has become the mostpredominant citrus-plant feeding swallowtail butterfly on the island. Asa result, it has become a significant pest of citrus plants, especiallynurseries. Young seedlings are particularly vulnerable, as they are lesscapable of surviving having their leaves eaten by butterfly larvaecompared to mature citrus plants. The butterflies prefer local speciesof citrus plants, such as lime plant and orange plant. Their consumptionof citrus plants is costly and wasteful, which makes them a tremendouspest to the citrus industry.

Measures taken by the industry to prevent attack on citrus plantnurseries thus far have proved expensive, ineffective, and in someinstances, deleterious to the environment. These methods includespraying, hand removal, and the use of physical barriers such asnetting.

SUMMARY

Methods of using a chemical attractant to control the feeding andbreeding behavior of target insects are disclosed. A host plant, whichgenerally contains an attractant for the target insect and is capable ofsupporting target insect offspring to control egg laying, is provided asubstantial distance from a plant of interest. The plant of interest isone in which target insects are naturally attracted to feeding orbreeding on in the wild, but is commercially or aesthetically valuableand therefore desirable of protecting from target insects that wouldharm it. No chemical attractant is applied to the plant of interest. Thehost plant and plant of interest may share a common chemical that actsas an attractant to the target insect. The chemical attractant isapplied to the host plant and induces target insects to lay eggs on it.The emerging juveniles feed on the host plant and, based on the tendencyof these insects to breed on the same type of plant on which they werereared, they lay the next generation of eggs on the same host plantspecies. The behavior of the target insect is controlled, for example,when a substantial number of mature offspring lay eggs on the same typeof host plant on which they were reared without repeat application ofthe chemical attractant.

In one aspect, the method includes inducing H. andraemon that are bredon lime plant to lay eggs on Piper amalago var. amalago plant. H.andraemon naturally prefer to breed on the type of plant on which theywere reared. Therefore, using this method results in an effectivecontrol measuring by which the egg-laying preference is maintained fromone generation to the next. By applying Piper amalago var. amalago plantoil, lime plant oil, limonene, or d-limonene in sufficient amounts toPiper amalago var. amalago plant, H. andraemon are shown to prefer tofeed and breed on Piper amalago var. amalago plant to lime plant.

In one aspect, a kit useful for controlling insect egg laying and/orfeeding is disclosed, which comprises a chemical attractant to attract atargeted insect. The chemical attractant can be selected from the groupconsisting of Piper amalago var. amalago plant oil, limonene,d-limonene, lime plant oil, or a combination thereof; an agriculturallyacceptable carrier; and instructions for applying the chemicalattractant to a host plant to induce egg laying or feeding on the hostplant instead of a plant of interest. The instructions direct that thehost plant be provided a particular distance from the plant of interest.In another aspect, the host plant is a Piper plant that containslimonene or d-limonene.

In any one of the above embodiments, providing the host plant consistsof planting enough of the host plant to support the egg-laying andfeeding of a target insect population.

In one or more embodiments, the behavior is controlled when 50% or moreof the offspring belonging to subsequent generations themselves matureand display a preference for laying eggs on the same type of host planton which they were reared without further application of the chemical.

In one or more embodiments, the application of the chemical attractantto the host plant is repeated as required to induce residualtarget-insect populations to lay eggs on the host plant.

In another aspect, a kit useful for controlling insect egg laying and/orfeeding is disclosed, and comprises a toxic extract for deterring targetinsects from laying eggs or feeding on a plant of interest. The toxicextract can be derived from any plant containing a compound that hastoxic effects on target insects. Specific examples of plants include,but are not limited to, Piper spp. such as Piper aduncum, Piper amalagovar. amalago, Piper hispidum, Piper fadyenii, and Piper nigrinodum.

In another aspect, a kit for repelling the feeding of target insectlarvae is disclosed. The kit provides a chemical attractant derived froma plant of interest. It also includes instructions for applying thechemical attractant to a host plant. Using this kit, gravid femaletarget insects are induced to lay eggs on the host plant, and becausethe target insects prefer to lay eggs on the type of plant on which theywere bred, the offspring of induce gravid females will overwhelminglyprefer to feed and breed on the type of host plant on which they werelaid.

In one or more aspects, the instructions direct the application of thechemical attractant to the host plant, or the extract to the plant ofinterest at least once daily, twice daily, weekly, twice weekly, or atleast once monthly.

In one or more of the above embodiments, the instructions directproviding enough of the host plant to support the egg-laying and feedingof a target insect population.

In any one of the above embodiments, the target insect is a species ofbutterfly.

In any one of the above embodiments, the target insect is a species ofcitrus swallowtail butterfly.

In any one of the above embodiments, the target insect is selected froma group consisting of Heraclides (syn. Papilio) andraemon Hubner,Heraclides andraemon: bonhotei Sharpe, Heraclides andraemon: andraemonHubner, Heraclides andraemon: hernandezi de la Torre, Heraclidesandraemon: tailori Rothschild & Jordan, Heraclides cresphontes Cramer,Heraclides hectorides Esper, Heraclides thoas L., Heraclides thoas:brasilensis Rothschild & Jordan, Heraclides thoas: melonius, Rothschild& Jordan, and Heraclides thersites, Fabricius.

In any one of the above embodiments, the chemical attractant isdetectable by the target insect.

In the embodiment in which the toxic extract is used as a repellant, thetoxic extract is detectable by the target insect.

In the embodiment in which the toxic extract is used as a pesticide, thetoxic extract may or may not be detectable by the target insect.

In one or more embodiments, the chemical attractant comprises oilderived from a host plant that contains limonene or d-limonene. Thechemical attractant can also be derived from a citrus plant.

In one or more of the embodiments, the chemical attractant comprises oilderived from the plant of interest, wherein the plant of interest isselected from the group consisting of Citrus aurantifolia (Christm.)Swingle (Rutaceae), Piper amalago var. amalago L. (Piperaceae), lime, C.limon (L.) Burm. F., lemon, C. sinensis (L.) Osbeck, sweet orange, C.reticulata Blanco, Mandarin orange, tangerine, C. paradisi Macf.,grapefruit, C. medica L., citron, C. aurantium L. Seville orange, C.grandis (L.) Osbeck, shaddock, pummelo, C. maxima (Burm.) Merr, ugli, C.reticulata Blanco×C. sinensis (L.) Osbeck, ortanique, C. mitis Blanco,calamondin, Fortunella margarita Lour. Swingle, kumquat (Rutaceae),Amyris P. Browne Rutaceae), Zanthoxylum L., Zanthoxylum martinicense(Lam.), Lantana camara, Jatropha podagrica, Canna indica, and Z. pterotaL. (Rutaceae).

In one or more of the embodiments, the plant of interest is a variety ofcitrus plant. The host plant can also be a Piper plant that containslimonene or d-limonene.

In any one of the above embodiments, the chemical attractant compriseslimonene or d-limonene.

In one or more of the above embodiments, the agent is an aerosol mixturecomprising the chemical attractant, the agriculturally acceptablecarrier, butane, and propane.

In one or more embodiments, the aerosol consists of butane, propane, andchemical attractant derived from limonene or d-limonene.

In one or more embodiments, the amount of chemical attractant in theaerosol mixture is about 0.01%-0.05%; 0.05%-0.10%, 0.15%-0.20%;0.20%-0.25%; 0.25%-0.30%; 0.30%-0.35%; 0.35%-0.40%; 0.40%-0.45%; or0.45%-0.50% by weight.

In any one of the above embodiments, the amount of limonene, d-limonene,Piper amalago var. amalago plant oil, lime plant oil, or a combinationthereof, is sufficient to attract the target insect.

In any one of the embodiments, the application consists of dipping,spraying, coating, diluting, covering, saturating, misting, fumigating,or dusting the host plant with the chemical attractant in an amountsufficient to affect the egg-laying behavior of the target insect.

In one aspect a pest control agent includes a chemical attractantselected from the group consisting of the Piper amalago var. amalagoplant oil, limonene, d-limonene, lime plant oil, and a combinationthereof; and an agriculturally acceptable carrier.

In one or more embodiments, the distance between the host plant andplant of interest is less than or equal to two kilometers.

In another aspect, target insect populations are controlled by applyinga toxic extract to plants of interest. The toxic extract causesdeformity or death when target insects ingest or come into physicalcontact with it. Toxic extract is derived from plants that contain toxinand no chemical attractant, or from host plants, which containattractant and a chemical that is toxic to target insects. The toxicextract can be used in at least two ways to control the behavior oftarget insect populations. In one embodiment, the toxic extract isapplied to a plant of interest and acts as a repellent by deterringtarget insects from feeding or breeding on the plant of interest. Inanother embodiment, the toxic extract is applied directly to eggs andlarvae of target insects as a pesticide to kill or deform targetinsects.

In any one of the above embodiments, the toxic extract is derived from aplant selected from the group consisting of Piper amalago var. amalagoL. (Piperaceae), Piper aduncum, Piper hispidum, Piper fadyenii, Cleomerutidosperma (Capperaceae), Pentas spp. (Rubiaceae), Lantana camara(Verbenaceae), Canna indica ((Cannaceae), Kalanchoe crassula(Crassulaceae), Pimenta dioica (Myrtaceae), Peperomia pellucida(Piperaceae), Phyllanthus amarus (Euphorbiaceae), Pilea microphylla:microphylla (Urticaceae), Oxalis corymbosa (Oxalidaceae), Begonia sp.(Begoniaceae), and Dracaena sandariana (Liliaceae) (Chinese bamboo).

In one or more embodiments, the toxin in the toxic extract is derivedfrom a Piper plant. The Piper plant can be, for example, Piper aduncum.

In another aspect, a pest control agent includes a toxic extract derivedfrom a plant containing a toxin, which causes death or deformity intarget insects when ingested an agriculturally acceptable carrier, andin some embodiments, limonene, or d-limonene.

In one embodiment of this aspect, the toxic plant is Piper aduncum.

One aspect is a method of controlling a target insect populationcomprising applying to a host plant an extract from a plant thatcontains a toxin, to deter target insects from laying eggs on the hostplant.

In one or more aspects, the toxin or agent is apiol.

In one or more aspects, the toxin is in the form of an emulsioncomprising apiol, ethanol, and an agriculturally acceptable carrier.

In one or more aspects, the amount of toxin in the emulsion is about0.1%-0.5%; 0.5%-1.0%, 1.0-1.5%, 1.5%-2.0%, 2.0%-2.5%, 2.5%-3.0%,3.0%-3.5%, 3.5%-4.0%, 4.0%-4.5%, 4.5%-5.0%, 5.0%-5.5%, 5.5%-6.0%,6.0%-6.5%, or 6.5%-7.0% by weight.

The advantages of using the disclosed methods, pest control agents, andkits are that they are useful natural and effective methods forcontrolling the feeding and breeding behavior of target insects.Specifically, the method, pest control agent, and kit are advantageousas natural and effective methods for controlling the feeding andbreeding behavior of citrus plant-feeding butterflies.

BRIEF DESCRIPTION OF THE DRAWINGS

It is to be understood that while the embodiments have been described inconjunction with the detailed description thereof, the foregoingdescription is intended to illustrate and not limit the scope of theembodiments, which is defined by the scope of the appended claims. Otheraspects, advantages, and modifications are within the scope of thefollowing claims.

FIG. 1 is an image of a gas chromatograph of lime plant essential oils.

FIG. 2 is an image of a gas chromatograph of Piper amalago var. amalagoplant essential oils.

FIG. 3 is an image of superimposed gas chromatographs of lime plant andPiper amalago var. amalago plant essential oils.

DEFINITIONS

For convenience, certain terms employed in the specification, examples,and appended claims are collected here. Unless defined otherwise, alltechnical and scientific terms used herein have the same meaning ascommonly understood by one of ordinary skill in the art to which thisinvention belongs. The initial definition provided for a group or termherein applies to that group or term throughout the presentspecification individually or as part of another group, unless otherwiseindicated.

The articles “a” and “an” are used herein to refer to one or to morethan one (i.e., to at least one) of the grammatical object of thearticle. By way of example, “an element” means one element or more thanone element.

The term “or” was used herein to mean, and was used interchangeablywith, the term “and/or,” unless context clearly indicates otherwise.

“Agriculturally acceptable carrier,” as that term is used herein, is amedium that is suitable for delivery of the population controlcomposition. The medium generally is compatible with active ingredientsand excipients of the population control composition and has a benigneffect on the environment. The excipient may be a wetting agent,spreading agent, deposit builder, adhesive, emulsifying agent,deflocculating agent, water modifier, or similar agent with or withouttoxic properties of its own. The excipient is intended to be used withthe active ingredient as an aid to its application or to its effect.

“Chemical attractant,” as that term is used herein, means the oils, suchas the essential oils, obtained from a plant of interest by chemical,physical, or mechanical means. The chemical attractant is known to beattractive to the target insect. It affects the feeding and breedingbehavior of target insects. Chemical attractants can be derived fromplants of interest or host plants.

“Host plant,” as that term is used herein, is a plant that is a hostplant for the target insect; it is naturally attractive to targetinsects. In some instances, the host plant is selected by the targetinsect for egg laying and the host plant serves as a source of food forthe emerging larvae. The host plant typically does not possesscommercial or aesthetic properties that impart value to the plant. It isused in the methods disclosed herein to protect plants possessingcommercial or aesthetic properties by serving as an alternative foodsource for target insects. The host plant may or may not be one that isnaturally attractive to the target insect, but in the case in which itis not naturally attractive, it can be made attractive by application ofcertain chemical attractants. In some instances, the host plant is not asource of food for the target insect. The host plant and plant ofinterest may or may not share a common chemical that acts as anattractant to the target insect.

“Target Insect,” as that term is used herein, is an insect that isnaturally attracted to the plant of interest and for which it is desiredto implement a measure of population control. Typically, the desire forpopulation control arises from the damage inflicted on the plant ofinterest by the target insect. The term ‘target insect’ encompasses theinsect at all stages of development and its offspring, including themature insect, eggs, pupae stage, and larvae.

“Pest control agent,” as that term is used herein, means an ingredientor a composition containing an ingredient that controls the populationof a target insect. Population control can be accomplished in a varietyof ways, including interruption of the feeding and/or breeding cycle ofthe target insect. In some embodiments, the pest control agent can serveas an attractant (for example, attracting the target insect away fromthe plant of interest or inducing female target insects to lay eggs onor consume plants treated with the attractant). In other embodiments,the pest control agent is used as a deterrent (for example, repellingthe target of interest from the plant of interest).

“Plant of interest,” as that term is used herein, is a plant to whichtarget insects are naturally attracted to feeding or breeding. The plantof interest has commercial or aesthetic properties that make it adesirable plant to protect from the harmful effects of feeding andbreeding target insects.

“Toxin,” as that term is used herein, is a chemical derived from a plantthat causes death or deformity in target insects that ingest or areotherwise exposed to the chemical. It also serves as a repellant (forexample, repelling the target of interest from the plant on which thetoxin is applied).

“Toxic extract,” as that term is used herein, means the oils, such asthe essential oils, obtained from a host plant that contains, inaddition to chemical attractant, compounds that deter target insectsfrom feeding or breeding on the plant to which the extract is applied.Toxic extract can also be derived from plants that contain toxicchemicals but no chemical attractant. The extract can be obtained bychemical, physical, or mechanical means. It is capable of causing deathor deformity in a target insect if ingested.

DETAILED DESCRIPTION

All publications, patent applications, patents, and other referencesmentioned herein are incorporated by reference in their entirety. Inaddition, the materials, methods, and examples are illustrative only andnot intended to be limiting. Unless otherwise defined, all technical andscientific terms used herein have the same meaning as commonlyunderstood by one of ordinary skill in the art to which this inventionbelongs. Although methods and materials similar or equivalent to thosedescribed herein can be used in the practice or testing of the presentinvention, suitable methods and materials are described below.

The methods and products described herein relate to the use of chemicalattractants and/or toxic extract to control the breeding behavior ofinsects, such as butterflies.

The methods involve using chemical attractants to control the egg-layingbehavior of target insects. A host plant is provided as a food sourcefor larvae, and is placed a particular distance from a plant of interestdesired to be protected from feeding target insects. The distance issmall enough to allow the host plant to be an alternate food sourceand/or breeding site for the target insects. However, the actualdistance will depend on the individual foraging patterns of particularinsect species, migration patterns of a particular insect species,ambient conditions, and the type of applicator used. In someembodiments, the host plants may be a distance of about 10 or morekilometers from the plant to be protected. When the target insect is abutterfly, and in particular a butterfly of the species H. andraemon,the distance between the host plant and plant of interest may be up toone to two kilometers apart.

Also disclosed is a pest control agent for attracting the feeding andbreeding of target insects. In this aspect, the pest control agentcomprises Piper amalago var. amalago plant oil, lime plant oil,limonene, d-limonene, or a combination thereof. It further comprises anagriculturally acceptable carrier. In one embodiment, the pest controlagent is an aerosol mixture, and is applied to the host plant byspraying the mixture onto the host plant.

A kit useful for controlling insect egg laying or feeding on a plant isalso disclosed. The kit contains chemical attractant in anagriculturally acceptable carrier and instructions for applying chemicalattractant to a host plant to induce the target insects to lay eggs orfeed on the host plant instead of nearby plants of interest.

In one or more embodiments, the chemical attractant is an extract fromthe host plant, which is known to be an attractant for the targetinsect. In other embodiments, the chemical attractant is a concentratedor purified component of the plant extract that is enriched in thechemical attractant. When the chemical attractant is applied to the hostplant in amounts greater than naturally occurring levels found in nearbyplants, the majority of gravid females prefer to lay eggs on host plantstreated with the chemical attractant to nearby untreated plants. Forsome target insects known to prefer to lay and breed on citrus plants,the chemical attractant contains limonene or d-limonene.

Using chemical attractant to control target insect feeding and egglaying is advantageous because it is a natural and effective method ofcontrolling the feeding and breeding behavior of insects such that thefeeding and breeding can be redirected from a valuable or aesthetic cropto a less valuable crop. Valuable and aesthetic crops are therebyallowed to thrive and are not consumed by target insects.

Certain insects display a natural tendency to prefer to breed on thesame type of plant on which they were reared. By inducing preferentialegg laying of target insects on the host plant instead of the plant tobe protected, subsequent generations exhibit a preference towardsfeeding and laying eggs on the host plant, thereby controlling the egglaying behavior of the target insect population. The behavior of theinsects is controlled when a substantial number, e.g., a majority, ofoffspring belonging to subsequent generations themselves mature anddisplay a preference for laying eggs on the same type of host plant onwhich it was reared without further application of the chemicalattractant. In one or more embodiments, more than 50% of the subsequentgeneration displays a preference for laying eggs on the same type ofhost plant on which they were reared. In one or more embodiments, morethan 55%, more than 60%, more than 65%, more than 70%, more than 75%,more than 80%, more than 85%, more than 90%, or more than 95% display apreference for laying eggs on the same type of host plant.

Chemical attractant is applied to the host plant to initially attracttarget insects and induce gravid females to lay eggs on it. When theoffspring emerge, they feed on the host plant on which they werehatched, and develop a preference for laying their eggs on thatparticular type of host plant. For the majority of target insects, thispreference for laying eggs on the same type of plant on which they werereared is maintained without reapplication of the chemical attractant.

In a particular embodiment, the target insects are a species ofbutterfly. In another embodiment, the target insects are gravid femalebutterflies. Particularly, the target insects are Citrus Swallowtailbutterflies, otherwise known as Heraclides (syn. Papilio) andraemon. Inanother embodiment, the target insects are Heraclides andraemon:bonhotei Sharpe, Heraclides andraemon: andraemon Hubner, Heraclidesandraemon: hernandezi de la Torre, Heraclides andraemon: tailoriRothschild & Jordan, Heraclides cresphontes Cramer, Heraclideshectorides Esper, Heraclides thoas L., Heraclides thoas: brasilensisRothschild & Jordan, Heraclides thoas: melonius, Rothschild & Jordan,and Heraclides thersites, Fabricius.

The host plant is used to provide a place for target insects to layeggs, and to provide food for larvae when the eggs hatch. Generally, thehost plant contains an attractant that causes target insects to feed orlay eggs on it. The host plant is also capable of supporting theoffspring of target insects. Any plant observed as a food source fortarget insects is an appropriate host plant. Suitable host plants havelittle or no commercial value, or are not susceptible to harm bybutterflies feeding on their leaves. The host plant can be one of, or acombination of, a variety of plants. For H. andraemon, this plantcontains the chemical attractant limonene or d-limonene. For instance,the host plant can be Piper amalago var. amalago plant, because H.andraemon has been observed to feed on Piper amalago var. amalago plantin the wild, and Piper amalago var. amalago plant contains d-limonene(see Table 1A). Citrus plants also contain d-limonene (see Table 1B). Ofcourse, because citrus plants also contain d-limonene and H. andraemonfeed on citrus plants in the wild, citrus plants can also be hostplants. Specific examples of host plants for H. andraemon include Piperamalago var. amalago L. (Piperaceae), Citrus aurantifolia (Christm.)Swingle (Rutaceae), lime, C. limon (L.) Burm. F., lemon, C. sinensis(L.) Osbeck, sweet orange, C. reticulata Blanco, Mandarin orange,tangerine, C. paradisi Macf., grapefruit, C. medica L., citron, C.aurantium L. Seville orange), Zanthoxylum L., and Zanthoxylummartinicense (Lam.).

The plant of interest is likely naturally attractive to target insects,typically is of commercial or aesthetic value in some respect, and istherefore in need of protection from feeding and breeding targetinsects. For the species of butterflies known to feed and lay eggs oncitrus plants, the plant of interest can be a plant from the citrusfamily. The plant of interest can be, for e.g., Citrus aurantifolia(Christm.) Swingle (Rutaceae), Piper amalago var. amalago L.(Piperaceae), lime, C. limon (L.) Burm. F., lemon, C. sinensis (L.)Osbeck, sweet orange, C. reticulata Blanco, Mandarin orange, tangerine,C. paradisi Macf., grapefruit, C. medica L., citron, C. aurantium L.Seville orange, C. grandis (L.) Osbeck, shaddock, pummelo, C. maxima(Burm.) Merr, ugli, C. reticulata Blanco×C. sinensis (L.) Osbeck,ortanique, C. mitis Blanco, calamondin, Fortunella margarita Lour.Swingle, kumquat (Rutaceae), Amyris P. Browne Rutaceae), Zanthoxylum L.,Zanthoxylum martinicense (Lam.), Lantana camara, Jatropha podagrica,Canna indica, or Z. pterota L. (Rutaceae). The host plant and the plantof interest can be the same or different species of plant. Thedifference is that the chemical attractant is applied to the host plantto induce egg laying and feeding on the host plant to protect the plantof interest from these insect activities.

The chemical attractant in the host plant can be the same chemicalattractant found in the plant of interest. For example, the host plantmay naturally produce the chemical attractant at lower levels than theplant of interest, which can cause target insects to be less attractedto the host plant in its natural state compared to plants of interestwith naturally higher levels of chemical attractant. Therefore, chemicalattractant is applied to host plants to make them more attractive tofeeding and egg-laying target insects, and to induce the insects to layeggs on it. For example, the chemical attractant applied to the hostplant can be derived from a plant containing limonene or d-limonene.More specifically, the chemical attractant can be derived from a citrusplant such as lime plant, Piper amalago var. amalago plant, or it can bean isolated component such as limonene or d-limonene.

According to one or more embodiments, host plant Piper amalago var.amalago can be treated with lime plant oil. Alternatively, Piper amalagovar. amalago plant can be treated with Piper amalago var. amalago plantoil or isolated limonene or d-limonene. The treated Piper amalago var.amalago plant is placed a distance from a plant of interest, e.g., limeplant. A mixture containing chemical attractant in the form of, e.g., anaerosol mixture, is applied to Piper amalago var. amalago plant in anamount sufficient to induce a number of target insects to feed or breedon Piper amalago var. amalago plant instead of lime plant. The distancebetween the Piper amalago var. amalago plant sprayed with, e.g., limeplant oil, and the plant of interest, is small enough to attract asubstantial number of target insects to feed or breed on the Piper plantinstead of the lime plant, but great enough to help prevent inadvertentapplication of the oil containing chemical attractant to the plant ofinterest. For example, the distance can be as great as 10 or morekilometers, or as small as two meters, depending upon foraging patternsof a particular insect, the migration distances of a particular insect,ambient conditions, and the type of applicator used.

The number of treated host plants should be sufficient to support thetarget population in the area.

H. andraemon is naturally attracted to feeding and breeding on limeplant and Piper amalago var. amalago plant. FIGS. 1 and 2 show images ofgas chromatographs for lime plant and Piper amalago var. amalago plants,respectively. Tables 1A and 1B list the compositions of Piper amalagovar. amalago plant oil and lime plant oil, respectively. Referring toFIG. 3, the chromatographs for Piper amalago var. amalago plant and limeplant superimposed revealed that a common oil, d-limonene, is found inboth lime plant and Piper amalago var. amalago plant Therefore, extractsderived from lime plant and Piper amalago var. amalago plant makeexcellent chemical attractants.

TABLE 1A Composition of Piper amalago var. amalago leaf oils by GasChromatography T_(R)/min Compound^(a) % Area RI^(b) ID^(c) 6.03 α-pinene4.67 932 GCMS, RI 6.81 β-pinene 6.52 1026 GCMS, RI 7.27 3-carene 9.421053 GCMS, RI 7.64 2,4-thujadiene 19.99 1075 GCMS, RI 8.02 d-limonene4.4 1097 S/M, RI 8.62 (R,S) linalool 5.93 1135 GCMS, RI 9.71 camphor0.29 1208 GCMS, RI 10.19 germacrene B 2.64 1251 GCMS, RI 10.25α-terpineol 1.75 1257 GCMS, RI 10.36 crypton 1.44 1267 GCMS, RI 11.18cuminaldehyde 0.32 1352 GCMS, RI 11.66 4-(1-methylethyl)benzenemethanol0.36 1408 GCMS, RI 12.3 piperitone 0.48 1494 GCMS, RI 13.07 α-cubene1.66 1610 GCMS, RI 13.19 1H-cyclopenta[1,3]cyclo- 0.78 1628 GCMS, RIpropa[1,2]benzene 13.76 caryophyllene 0.87 1712 GCMS, RI 14.35γ-murolene 1.04 1788 GCMS, RI 14.45 α-gurjunene 1.64 1801 GCMS, RI 14.5β-guainene 0.74 1807 GCMS, RI 14.64 β-cadinene 0.56 1824 GCMS, RI 14.71β-gurjunene 2.94 1832 GCMS, RI 14.89 α-cadinene 1.84 1853 GCMS, RI 14.98calamenene 4.01 1864 GCMS, RI 15.21 nerolidol 1.16 1891 GCMS, RI 15.79spathulenol 1.23 1960 GCMS, RI 16.03 caryophyllene oxide 1.17 1988 GCMS,RI 16.33 calarene 1.64 2022 GCMS, RI 16.44 α-guaiene 0.78 2035 GCMS, RI16.5 copaene 2.15 2042 GCMS, RI 16.66 δ-selinene 1.33 2060 GCMS, RITotal 83.75 ^(a)Elution order on HP capillary column. ^(b)Retentionindex relative to n-alkane series(C₅-C₃₀ excluding C₂₇ and C₂₉) on HPDB-5 column, ^(c)GCMS identification by Gas-chromatography-Massspectroscopy, *matched by authentic internal standard

TABLE 1B Composition of Lime Plant Essential Oils Obtained by GasChromatography T_(R)/min Compound^(a) % Area RI ^(b) ID^(c) 4.47 4hydroxy-4-methyl-2-pentanone 10.13 994 GCMS, RI 6.92 decane 1.9 1002GCMS, RI 7.59 d-limonene 22.04 1209 GCMS, RI 8.57 undecane 3.49 1095GCMS, RI 9.54 citronellal 1.22 1484 GCMS, RI 9.93 thujone 1.92 1418GCMS, RI 10.14 dodecane 0.54 1194 GCMS, RI 10.62 nerol 8.62 1752 GCMS,RI 10.93 geraniol 1.22 1797 GCMS, RI 11 neral 9.13 1701 GCMS, RI 11.40geranial 12.91 1739 GCMS, RI 12.18 geranyl acetate, (Z) 2.9 1727 GCMS,RI 12.77 geranyl acetate (E) 2.74 1773 GCMS, RI 17.73 squalene 18.532633 GCMS, RI Total 97.28 ^(a)Elution order on HP capillary column. ^(b)Retention index relative to n-alkane series(C₅-C₃₀ excluding C₂₇ andC₂₉) on HP DB-5 column, ^(c)GCMS identification byGas-chromatography-Mass spectroscopy

The amount and frequency for applying the chemical attractant to thehost plant can be repeated as required to increase the percentage of theinsect population that initially is attracted to the host plant species,or to induce residual insect populations to lay eggs on the host plant.Residual insect populations consist of mature target insects that preferto feed and/or breed on plants other than that on which they werereared.

The chemical attractant is applied in an amount sufficient to inducetarget insects to feed or breed on the host plant. The amount sufficientto attract the target insects varies depending on the manner in whichthe chemical attractant is applied to the host plant and the type ofhost plant to which the chemical attractant is applied. Once applied,the chemical attractant lasts for about 12 to 24 hours or more,depending on the chemical stability and volatility of the chemicalattractant and ambient conditions.

Further, the chemical attractant is applied to the host plant in anamount and a number of times sufficient to be recognized by targetinsect and be attractive to them. In one embodiment, the chemicalattractant is applied to the host plant once per day. The chemicalattractant can be applied to the host plant two, three, four, five, ormore times per day. In another embodiment, the chemical attractant isapplied to the host plant once per week. In other embodiments, thechemical attractant is applied to the host plant twice per day. In otherembodiments the chemical attractant is applied to the host plant once,twice, three or more times per month. It can be applied before arainfall, or after a rainfall. The chemical attractant can be appliedwhen new leaves grow on the host plant. In another embodiment thechemical attractant is applied to the host plant when new leaves grow onthe plant of interest.

The amount of chemical attractant used varies depending on the manner inwhich it is applied and the type of host plant to which it is applied.The amounts also vary depending on the species of target insect andambient conditions. Using Piper amalago var. amalago plant oil in analkane hydrocarbon aerosol mixture as an example, generally, the amountof Piper amalago var. amalago plant oil in the aerosol mixture requiredto induce a number of target insects to feed or breed on Piper amalagovar. amalago plant instead of lime plant is about 0.01%-0.50% by weight.For example, the amount of Piper amalago var. amalago plant oil in theaerosol mixture is about 0.01%-0.05%; 0.05%-0.10%, 0.15%-0.20%;0.20%-0.25%; 0.25%-0.30%; 0.30%-0.35%; 0.35%-0.40%; 0.40%-0.45%; or0.45%-0.50% by weight.

In another aspect, the population size of target insects can becontrolled by applying toxic extract to a plant of interest. Plants ofinterest naturally contain chemical attractant that induce targetinsects to breed on the plants of interest. However, toxic extractsderived from plants that may or may not also contain a chemicalattractant or attractants, are poisonous to target insects and repelthem. Therefore, in the embodiment in which toxic extracts are appliedto the leaves of plants of interest, gravid female target insects aredeterred from laying eggs or feeding on the treated plants. In anotherembodiment, toxic extracts are used as a pesticide and are applieddirectly to target insects. As a result, the majority of the offspringeither die in early stages of the life cycle, or become deformed adultsthat are unable to fly and thus unable to reproduce and functionnormally. Thus, these methods thereby reduce the population size of, andprotect plants of interest from being destroyed by feeding targetinsects.

In another embodiment, a kit useful for controlling insect egg laying orfeeding on a plant is disclosed in which the kit contains toxic extractin an agriculturally acceptable carrier. Also included are instructionsfor applying the toxic extract to a plants of interest to repel targetinsects from laying eggs or feeding on treated plants of interest.

The toxic extract can be derived from a plant containing any toxin thatrepels, kills, or deforms target insects. For example, the toxic extractcan be derived from Piper aduncum, Piper hispidum, Piper fadyenii, Piperamalago var. amalago plant, or it can be an isolated component such asapiol, which is a pesticidal compound found in Piper aduncum plant (seeTable 1D).

In one embodiment of this aspect, toxic extract, derived from, forexample, Piper aduncum, can be used to treat plants of interest toprotect them from feeding and breeding H. andraemon. Treatment isconsidered effective when, e.g., a majority of target insects refuse tofeed or breed on the plant to which the toxic extract is applied. In oneor more embodiments, more than 50% of the target insects are deterredfrom feeding or breeding on the plant to which the extract is applied.In one or more embodiments, more than 55%, more than 60%, more than 65%,more than 70%, more than 75%, more than 80%, more than 85%, more than90%, or more than 95% of the target insects are deterred from feeding orbreeding on the plant to which the extract is applied.

In the aspect in which the toxic extract is applied to the plant ofinterest to repel target insects from feeding or breeding on plants ofinterest, it is applied in an amount and a number of times sufficient tobe recognized by target insect and be deter them from feeding orbreeding on the plant of interest. In the aspect in which the toxicextract is applied as a pesticide to eggs laid or larvae feeding onplants of interest, the toxic extract is applied in an amount and numberof times sufficient to kill or deform target insects. In one embodimentof these aspects, the toxic extract is applied to the plant of interestonce per day. In another embodiment, it is applied to the plant ofinterest two, three, four, five, or more times per day. In anotherembodiment, the toxic extract is applied to the plant of interest onceper week. In other embodiments, the toxic extract is applied to theplant of interest twice per day. In other embodiments, the toxic extractis applied to the plant of interest once, twice, three or more times permonth. It can be applied before a rainfall, or after a rainfall. Thetoxic extract can be applied when new leaves grow on the plant ofinterest. In another embodiment, the toxic extract is applied to theplant of interest when new leaves grow on the plant of interest.

The toxic extract is applied in an amount sufficient to deter targetinsects from feeding or breeding on the host plant to which the toxicextract is applied. The amount sufficient to deter the target insectsvaries depending on the manner in which it is applied to the host plantand the type of host plant. Once applied, the toxic extracts last for 12to 24 hours, depending on ambient conditions and the volatility of thetoxic extract. Application to plants of interest can be repeated asrequired to continue to deter target insects from feeding and layingeggs on treated plants.

The amount of toxic extract used varies depending on the manner in whichit is applied and the type of plant of interest to which it is applied.The amounts also vary depending on the species of target insect andambient conditions. Using Piper aduncum plant oil in an ethanol orwater-based emulsion as an example, generally, the amount of Piperaduncum plant oil in the mixture required to repel the feeding andbreeding of H. andraemon on plants of interest is about 0.1%-0.5%;0.5%-1.0%, 1.0-1.5%, 1.5%-2.0%, 2.0%-2.5%, 2.5%-3.0%, 3.0%-3.5%,3.5%-4.0%, 4.0%-4.5%, 4.5%-5.0%, 5.0%-5.5%, 5.5%-6.0%, 6.0%-6.5%, or6.5%-7.0% by weight, preferably 5%. When used as a pesticide to kill ordeform target insects on contact, the amount of Piper aduncum plant oilin the mixture required is about 0.1%-0.5%; 0.5%-1.0%, 1.0-1.5%,1.5%-2.0%, 2.0%-2.5%, 2.5%-3.0%, 3.0%-3.5%, 3.5%-4.0%, 4.0%-4.5%,4.5%-5.0%, 5.0%-5.5%, 5.5%-6.0%, 6.0%-6.5%, or 6.5%-7.0% by weight,preferably 2%.

Table 1C shows the chemical composition of the oil derived from Piperamalago var. nigrinodum plant. H. andraemon is not known to feed onPiper amalago var. nigrinodum plant in the wild. This is consistent withthe observed unsuitability of Piper amalago var. nigrinodum plant to bea host plant for H. andraemon, and is further confirmation thatd-limonene is a chemical attractant for H. andraemon. The oil recoveredis very pale yellow to colorless with an odor similar to that of blackpepper. The leaves or extract from Piper amalago var. nigrinodum plant,as shown in Table 1C, do not contain the attractant d-limonene.

As shown in Table 1D, the oil derived from Piper aduncum plant also doesnot contain the attractant d-limonene, but does contain a significantamount of the oil of the pesticide apiol (71.06%), which makes Piperaduncum as excellent deterrent. H. andraemon is not known to feed onthis species of plant in the wild.

TABLE 1C Composition of Piper amalago var. nigrinodum Plant EssentialOils Obtained by Gas Chromatography T_(R)/min Compound^(a) % Area RI^(b)ID^(c) 4.46 heptane 10.6 700 GCMS, RI 10.95 α-pinene 10.95 1032 GCMS, RI6.31 camphene 1.77 1076 GCMS, RI 6.64 sabinene 10.42 1158 GCMS, RI 6.81β-pinene 11.02 1124 GCMS, RI 7.28 3-carene, 1.71 1130 GCMS, RI 7.62β-phellandrene 16.25 1279 GCMS, RI 8.61 (R,S) linalool 2.66 1151 GCMS,RI 10.25 α-terpineol 1.62 1688 GCMS, RI 10.36 crypton 1.8 1124 GCMS, RI11.17 4-(1-methylethyl)benzenaldehyde 0.46 1789 GCMS, RI 11.654-(1-methylethyl) benzenemethanol 0.29 1517 GCMS, RI 11.69 bornylacetate 0.66 1560 GCMS, RI 12.55 (+)-2-carene 0.35 1110 GCMS, RI 13.07copaene, 0.48 1376 GCMS, RI 13.77 caryophyllene 1.12 1707 GCMS, RI 14.01valencene 0.51 1714 GCMS, RI 14.98 calamenene 1.23 1875 GCMS, RI 15.21trans -nerolidol 1.77 2058 GCMS, RI 15.79 spathulenol, 2.56 2126 GCMS,RI 16.03 caryophyllene oxide 1.51 1973 GCMS, RI 17.18 β-gurjunene 7 1611GCMS, RI 17.27 α-guaiene 9.94 1657 GCMS, RI Total 96.68 ^(a)Elutionorder on HP capillary column. ^(b)Retention index relative to n-alkaneseries(C₅-C₃₀ excluding C₂₇ and C₂₉) on HP DB-5 column, ^(c)GCMSidentification by Gas-chromatography-Mass spectroscopy

TABLE 1D Composition of Pipe aduncum Leaf Essential Oils Obtained by GasChromatography Retention Compound^(a) time RI Area 1 α-pinene 6.71 9530.31 2 β-pinene 7.5 979 0.34 3 β-myrcene 7.66 988 0.14 4 m-cymene 8.31026 0.87 5 trans-β-ocimene 8.46 1035 1.98 6 cis-β-ocimene 8.64 10462.97 7 3-carene 8.87 1060 0.54 8 α-terpinolene 9.32 1087 0.22 9terpinen-4-ol 10.92 1182 1.61 10 d-piperitone 12.27 1260 3.96 11 copaene14.27 1382 0.42 12 germacrene A 14.45 1394 0.26 13 caryophyllene 14.971430 1.68 14 α-caryophyllene 15.48 1466 0.65 15 β-cubene 15.83 1490 1.3016 myristicin 16.31 1526 3.53 17 elemicin, (Z) 16.59 1547 0.41 18nerolidol 16.77 1561 0.28 19 γ-elemene 16.9 1572 0.23 20 (−)-spathulenol17.11 1588 0.23 21 caryophyllene oxide 17.2 1595 0.86 22 γ-selinene17.35 1607 0.79 23 apiol 17.65 1632 71.06 244,5-dihydro-1,3-diphenyl-1H-pyrazole, 17.77 1642 0.37 25 calarene 17.921655 0.55 Total 95.56 ^(a)Elution order on HP capillary column.^(b)Retention index relative to n-alkane series(C₅-C₃₀ excluding C₂₇ andC₂₉) on HP DB-5 column, ^(c)GCMS identification byGas-chromatography-Mass spectroscopy

Chemical attractants can be comprised of one of several chemicals. Inone embodiment, the chemical attractant is derived from host plants,which contain attractants and toxins. In another embodiment, thechemical attractant is derived from plants of interest that containattractants but not toxins. The chemical attractant can be an extract ofsoluble components from either the host plant or plant of interest.

Similarly, toxic extracts can be comprised of one of several chemicals.In one embodiment, the toxic extract is derived from plants that containtoxins. In another embodiment, the toxic extract is derived from hostplant, which contain, in addition to chemical attractant, toxins thatrepel target insects. Further, the toxic extract can be an extract ofsoluble components from a host plant, or other type of plant thatcontains a chemical that repels target insects.

Chemical attractants and toxic extracts can contain components solublein hydrophobic medium as an oil extract or in hydrophilic and aqueousmedia as a water extract. Extracts can be obtained using techniquesknown in the art, such as hydrodistillation, hydro diffusion, coldpressing, extraction using a hydrocarbon solvent, super critical fluidextraction using carbon dioxide and other super critical fluids, steamdistillation, fractional distillation, enfleurage extraction, macerationextraction, including those processes with zeolite removal of the waterafter maceration, and spinning cone extraction. Isolation andpurification of individual or groups of chemical compounds can beaccomplished using methods established in the art, such as gas, liquid,including such developments as high performance liquid chromatography(HPLC), high performance liquid chromatography guided by massspectroscopy (HPLC-MS), high performance liquid chromatography guided bynuclear magnetic resonance spectroscopy (HPLC-NMR), multiple highperformance liquid chromatography (HPLC-HPLC), ultra-performance liquidchromatography (UPLC), and ultra-performance liquid chromatographycoupled to-time-of flight mass spectrometry (UPLC-TOF-MS),countercurrent or centrifugal partition chromatography (CCC or CPC),column, and size-exclusion chromatography, homogenization, distillation,and fractional distillation.

Chemical attractant and toxic extract can be applied to plants, and inthe case of toxic extract, directly to target insects, in the form of aspray, paste, gum, oil, solution, aerosol, mist, dust, fume, and/or gas.Chemical attractant is applied in an amount sufficient to affect theegg-laying behavior of target insects. The egg-laying behavior isaffected in that the plant on which the target insect prefers to feed orbreed changes from the plant of interest (its food source and preferredplace for egg laying under natural conditions) to the particular type ofhost plant on which the target insect was reared. The chemicalattractant can be in the form of an aerosol, where the aerosol is amixture of butane, propane, and the chemical attractant, and can beapplied by spray or mist. Toxic extract is applied in an amountsufficient to deter target insects from feeding or breeding in treatedplants of interest. Toxic extract can be in the form of an emulsion withwater and a suitable alcohol, such as ethanol.

Species of plant were collected and deposited in the Herbarium in theDepartment of Life Sciences at the University of West Indies, Mona,Jamaica. For identification purposes, the plants used in these methodswere assigned the following accession numbers:

Plants Accession Numbers Citrus aurantifolia 35266 Citrus sinensis 35267Piper amalago var. amalago 35268 Piper amalago var. nigrinodum 35291Zanthoxylum martinicense 35289 Piper aduncum 35435

EXAMPLES Example 1

To rear H. andraemon larvae on citrus plants and Piper amalago var.amalago plants, 77 larvae were laid on various citrus plant species,including lime plant, Citrus sinensis (sweet orange), and Citrusreticulata (Mandarin orange). The larvae were then manually removed andreared on Piper amalago var. amalago plants. The larvae were transferredat different stages, but early in their life cycles (first, second, orthird instars). Measurements of the widths of the broadest part of thepupae were measured, and time to pupation was taken to determine hostplant suitability. The larvae were placed in a cage of about 3 meters by3 meters by 2.5 meters and were exposed to normal ambient environmentalconditions. Statistical analysis was used to determine whether therewere significant differences between the size of the larvae bred onPiper amalago var. amalago plant and citrus plants.

Time to pupation was determined by noting the time taken for larvae topupate after hatching for both lime plant and Piper amalago var. amalagoplant-reared larvae. The leaves of both plants on which the eggs werelaid were removed from the cage and replaced with fresh leaves. Thenumber of pupae and number of days the larvae took to pupate wasrecorded. Statistical analysis was used to determine whether there weresignificant differences between the times it took larvae bred on limeplant and larvae bred on Piper amalago var. amalago plant to pupate.

Various measurements were taken to compare the suitability of Piperamalago var. amalago plant as a host-plant substitute for citrus plants.Once mature, the feeding and breeding behavior of Piper amalago var.amalago plant-reared butterflies was compared to lime plant-rearedbutterflies. The experiments and the results, which demonstrated thatthe feeding and breeding behavior of butterflies can effectively becontrolled with the use of chemical attractants, are summarized in theExamples 2-8.

Example 2

Fresh whole leaves of lime plant and Piper amalago var. amalago plantwere used to extract oil for application to host plants. The leaves werethen hydrodistilled for up to 4 hours using a Clevenger-type apparatus.The oils were collected during distillation at one hour intervals, andwere dried over anhydrous sodium sulfate to yield substantially clearoils. Extractions were done in triplicate and the average values wereused. The oils were weighed and stored at 5° C.

Hydrodistilling 952.7 g of lime plant yielded 1.44 g of oil, 0.151% byweight; 378.6 g of Piper plant yielded 0.446 g of oil, 0.085% by weight;193.2 g Citrus sinensis leaves yielded 0.782 g of oil, 0.405% by weight;794 g of P. amalago, var. nigrinodum leaves yielded 0.719 g of oil,0.091% by weight; and 207.9 g of Zanthoxylum martinicense leaves yielded0.0365 g of oil, 0.0176% by weight.

A Varian CP-3800 gas chromatograph interfaced with a Flame ionizationdetector (FID) was used to analyze the extracted oils. The gaschromatograph was equipped with a WCOT fused silica coated with CP WAX52CB capillary column (length 60 m×inner diameters 0.25 mm; 0.25 μm filmthickness). The carrier gas was Nitrogen, at a flow rate of 1 mL min⁻¹,split 1:100. The injector temperature was 250° C. The column oventemperature of 40° C. was held for 1 minute, then increased from 40° C.to 100° C., at a rate of 10° C. min⁻¹ held for 1 minute, then increasedfrom 100° C. to 200° C. at a rate of 20° C. min⁻¹, held for 1 minute,and finally increased from 200° C. to 250° C. at a rate of 10° C. min⁻¹,and held for 25 minutes. The FID temperature was maintained at 300° C.

The retention indices (RI) were calculated using a mixture of homologousseries of n-alkanes C₈-C₂₅ analyzed under the same conditions as eachoil sample (GC-FID analysis). RI values were calculated using Kovats'procedure for temperature programming GC equations. (IUPAC. Compendiumof Chemical Terminology, Kovats (Retention) Index; 1997).

The chemical compositions of the oils were also determined using GC-MSusing a Hewlett Packard (HP) 6890 system Gas Chromatograph interfacedwith a HP-5973 Mass Spectrometer. The gas chromatograph was equippedwith a DB-VRX fused silica column (length 20 m×internal diameters 0.18mm, film thickness of 1 μm). Analytical conditions employed were Heliumfor a carrier gas at a flow rate of 1 mL min⁻¹, split less mode, with aninjector temperature of 250° C., interface temperature of 280° C. Thetemperature program was the same used for the GC-FID analyses previouslydescribed. The mass spectra data were collected with ionization energyof 70 eV and a mass range of 50-500 M/Z. An n-alkane mixture was alsoanalyzed under the same temperature program and other conditions, andthe Retention Indices (RI) calculated for each compound.

The components in the oils were matched with mass spectral data of theNIST 98 library. Peaks that had a peak quality match greater than 70%were considered matches with the compound from the library. Peaks ofsubstantial quantity but poor quality were identified either by matchinggas chromatography analyses with authentic compounds run in severalprograms, or by a comparison of the retention indices against thepublished data in Adams. (Adams, Robert, Identification of oilcomponents by gas chromatography/quadruple mass spectrometry; 2001:9-40). The presence of d-limonene in Piper amalago var. amalago leaf oilwas confirmed by matching against authentic d-limonene in severaltemperature programs: the standard program mentioned previously, and twoadditional temperature programs, firstly, heated initially to 40° C. andheld at this temperature for 3 min., then heated from 40° C. to 80° C.at a rate of 5° C. min⁻¹, and held at this temperature for 1 min., thenheated from 80° C. to 200° C. at a rate of 10° C. min⁻¹, and held atthis temperature for 2 min., then finally heated from 200° C. to 250° C.at a rate of 10° C. min⁻¹, and held at 250° C. for 10 min., andsecondly, heated initially to 40° C. and held at this temperature for 3min., then heated from 40° C. to 120° C. at a rate of 10° C. min⁻¹ andheld at 120° C. for 3 min., then heated from 120° C. to 180° C. at arate of 20° C. min⁻¹ held at that temperature for 5 min., then finallyheated from 180° C. to 250° C. at a rate of 20° C. min⁻¹, and held atthat temperature for 1 min.

Fresh leaves were collected to extract the oils for analysis by gaschromatography. The leaves were weighed and hydrodistilled for up tofour hours, using a Clevenger type apparatus. The extractions were donein triplicate and the essential oils were collected during distillationat hourly intervals. The oils obtained were then dried over anhydroussodium sulfate and yielded clean, clear oils. The oils were then weighedand stored at 5° C., in a refrigerator for further analysis.

The aerosol mixture for application to citrus plant and Piper amalagovar. amalago plant materials was prepared by combining lime plant oil,Piper amalago var. amalago plant oil, d-limonene, or a combinationthereof, with an alkane hydrocarbon mixture consisting of 28% butane and72% propane. The aerosol was filled to a concentration of, e.g., 0.25%by weight of Piper amalago var. amalago plant oil in the hydrocarbonmixture or, e.g., 0.45% by weight of lime plant oil in the hydrocarbonmixture.

Example 3

To determine the suitability of Piper amalago var. amalago plant as ahost plant, the size of larvae were compared after feeding on eitherPiper amalago var. amalago plant or lime plant. In this experiment,larvae were reared on Piper amalago var. amalago plant or lime plant tocompare the size of the larvae after they fed on one of either plant.

TABLE 2 Comparison of pupae size (mm) Pupae bred on Piper amalago Pupaebred on var. amalago plant citrus plant Population size: 77     72    Mean width: 8.710 mm. 8.853 mm. Standard Deviation: 1.0354 1.0978

The results, reflected in Table 2, show that the difference in the meanwidths of pupae bred on Piper plant and lime plant was minimal at about0.1 mm. The Levene's test for equality of variance indicated that thevariance in size were not significantly different (p=0.432). Because nosignificant difference was found between the size of pupae bred on Piperamalago var. amalago plant versus lime plant, Piper amalago var. amalagoplant was determined to be a suitable host plant substitute for limeplant.

Example 4

The sex ratios of the larvae were compared to determine the suitabilityof Piper amalago var. amalago plant as a host-plant substitute for limeplant. To determine the sex ratios of butterflies bred on lime plant andPiper amalago var. amalago plant, a random population of 45 lime plantand 39 Piper amalago var. amalago plant-reared butterflies was selected.The butterflies were reared to adulthood on either lime plant or Piperamalago var. amalago plant.

TABLE 3 Comparison of sex ratios of H. andraemon larvae bred on Piperamalago var. amalago plant and citrus plant Male Female Totals Observedcitrus-plant 24 21 45 reared: Observed Piper-plant 17 22 39 reared:Totals 41 43 84

TABLE 3A Chi-squared values observed, and expected number of male andfemale butterflies from citrus-reared larvae Male Female Totals Observed(O) 24 21 45 Expected (E) 22.5 22.5 45 χ² 0.1977 Yates Corrected value0.0889

TABLE 3B Chi-squared values observed, and expected number of male andfemale butterflies from P. amalago var. amalago -reared larvae MaleFemale Totals Observed (O) 17 22 39 Expected (E) 19.5 19.5 39 χ² 0.641Yates Corrected value 0.410

TABLE 3C Sex ratios for Piper amalago var. amalago plant-reared andcitrus-plant reared larvae Male Female Totals citrus plant reared: 24 2145 Piper amalago var. amalago 17 22 39 plant reared: Totals 41 43 84

TABLE 3D Calculated data for chi-squared contingency test on sex ratiosfor Piper amalago var. amalago plant-reared and citrus-reared larvaeMale Female Totals Observed citrus plant reared 24 21 45 (O) Expected(E) 21.96 23.04 45 Observed Piper amalago 17 22 39 var. amalago plantreared (O) Expected (E) 19.04 19.96 39 χ² 0.796

After the emergence of the adult butterfly, sex was determined byviewing the pupal shell under a light microscope at 10× magnification.The shells were noted as either male or female based on the appearanceof two spots in either the first or second segment, according to themethod by George Warenecke, The Young Specialist Looks at Butterfliesand Moths; 1964: 34-35.

The values were assigned a chi square value of 0.796 compared to the Pvalue at 0.001 of 10.10.83 (Table 3D). The calculated chi squared valueswere less than the tabulated values (p>0.001), which were in agreementthat the observed data does not differ significantly from the expecteddata. Variation in the sex ratios was attributed to chance, and thenumber of male and female butterflies obtained from each host plant wasin accordance with that which is expected from natural selection, for asex ratio of 1:1.

The results of this experiment indicate that Piper amalago var. amalagoplant is a suitable host-plant substitute for lime plant.

Example 5

The suitability of Piper amalago var. amalago plant as a host plant wasfurther experimented by comparing life cycles of H. andraemon bred onPiper amalago var. amalago plant to those bred on lime plant. The lifecycles of H. andraemon bred on Piper amalago var. amalago plant and H.andraemon bred on lime plant were very similar. The days from hatchingto pupation were about 17 to 23 days, and the emergence of the adultfrom the pupa was about 10 days to 3 weeks.

Example 6

To further evaluate the suitability of Piper amalago var. amalago plantto serve as a host plant in place of naturally preferred citrus plant,the number of offspring yielded from egg laying on Piper amalago var.amalago plant was compared to that from citrus plants.

TABLE 4 Number of eggs laid on Piper amalago var. amalago plant andcitrus plant by H. andraemon reared on Piper amalago var. amalago andcitrus plants Number of eggs Plant on which Number of laid on PiperButterflies eggs laid on amalago var. were reared citrus plant amalagoplant Total Larvae reared on 64 (89%)  8 (11%) 72 (100%) citrus plant(%) Larvae reared on Piper 2 (4%) 53 (96%) 55 (100%) amalago var.amalago (%) Total no. of eggs 66 61 127

TABLE 4A Number of eggs laid by Piper amalago var. amalago plant-rearedbutterflies on individual host plants Number of eggs laid on PiperNumber of amalago var. eggs laid on Total eggs laid amalago plant citrusplant 55 53 2 Percentage of total eggs 96% 4% laid

TABLE 4B Number of eggs laid by citrus-plant bred butterflies onindividual host plants Number of eggs laid on Piper Number of amalagovar. eggs laid on Total eggs laid amalago plant citrus plant 72 8 64Percentage of total eggs 11% 89% laid

TABLE 4C Observed and expected number of eggs laid on each host plant byH. andraemon Number of eggs Number of laid on Piper eggs laid on amalagovar. citrus plant amalago plant Total Larvae reared on 64 * (37.417) 8 * (34.583) 72 citrus plant Larvae reared on Piper  2 * (28.583) 53 *(26.417) 55 amalago var. amalago Total 66 61 127 * values in bracketsare calculated expected values Calculated chi-square value = 90.79, P at0.001 at 1 degree of freedom = 10.83.

Table 4 shows the startling result that gravid females lay eggs on thesame species of host plant on which they were reared. In this example,Piper amalago var. amalago plant-bred butterflies were allowed to choosebetween laying eggs on Piper amalago var. amalago plant and lime plant.The plants were arranged in varying directions to avoid selection basedon orientation. A chi-squared contingency test on the relationshipbetween the plant species of rearing and the plant species of egg-layinggave a highly significant result, (X²=90.79 at 1 degree of freedom,giving a value of p<0.001), indicating that the species that thebutterflies are reared on, does influence the species on which theyprefer to lay their eggs.

Referring to Table 4A, in a population of 55 eggs laid by Piper amalagovar. amalago plant-bred butterflies, when given the choice of layingeggs on Piper amalago var. amalago plant or lime plant, 53 eggs, or 96%,were laid on Piper amalago var. amalago plant, and only 2 eggs, or 4%,were laid on lime plant. Similarly, lime plant-reared butterflies weregiven the choice of laying eggs on Piper amalago var. amalago and limeplants. Table 4B shows that 64 eggs, or 89%, were laid on lime plant bylime plant-reared butterflies, and only 8 eggs, or 11%, were laid onPiper amalago var. amalago plant.

Table 4C shows that the calculated chi square value was 90.79, which wasgreater than the tabulated value of 10.83 at P=0.001. The eggs laid onthe host plant other than the species on which the larvae werethemselves reared were attributed to chance.

These results show that the butterflies overwhelmingly preferred to reartheir offspring on the same type of plant on which they were reared.Therefore, butterflies initially induced by a chemical attractant to layeggs on Piper amalago var. amalago plant instead of citrus plant yieldoffspring that also prefer that species of plant.

Example 7

The faithfulness of egg laying of lime plant-bred butterflies on limeplant versus Piper amalago var. amalago plant was compared to furtherdetermine the suitability of Piper amalago var. amalago plant as a limeplant substitute for controlling feeding and egg laying behaviors ofbutterflies.

H. andraemon larvae were reared on lime plant. When they pupated andwere ready to lay eggs, they were given the choice of plants on which tolay eggs: unsprayed lime plants, unsprayed Piper amalago var. amalagoplants, Piper amalago var. amalago plants sprayed with lime plant oil,and Piper amalago var. amalago plants sprayed with d-limonene. Egglaying was permitted to continue until the butterflies ceased layingeggs. Nearly 100% of the eggs laid hatched into larvae.

Table 5 summarizes the results of an experiment in which thefaithfulness of egg laying on the species of plant on which thebutterflies were reared was confirmed. In this experiment, both limeplant oil and d-limonene were sprayed onto Piper amalago var. amalagoplant to determine whether the butterflies discriminated between thetwo.

TABLE 5 Egg laying of citrus-plant bred H. andraemon given a choice ofplants on which to lay No. of Plants eggs laid % a. Unsprayed lime plant93 44.71 b. Unsprayed Piper amalago var. amalago 11 5.29 c. Piperamalago var. amalago sprayed with 52 25.00 lime plant oil d. Piperamalago var. amalago sprayed with 52 25.00 d-limonene Totals 208 100.00

As expected, most of the eggs were laid on unsprayed lime plants,confirming that lime plant-reared butterflies preferentially re-lay eggson lime plants. Only 5.29% of eggs were laid on unsprayed Piper amalagovar. amalago plants. The surprising result, which is not naturallyoccurring, was that gravid females perceived Piper amalago var. amalagoplants sprayed with either lime plant oil or d-limonene similarly, asreflected by the 52 eggs laid on Piper amalago var. amalago plantssprayed with lime plant oil and the 52 eggs laid on Piper amalago var.amalago plants sprayed with d-limonene. Moreover, these results showthat sprayed Piper amalago var. amalago plants, on which 50% of eggswere laid, were perceived similar to unsprayed lime plants, and aretherefore suitable for inducing egg laying on species other than citrusplant.

Example 8

The faithfulness of egg laying of Piper amalago var. amalago plant-bredbutterflies on Piper amalago var. amalago plant versus lime plant wascompared to further determine the suitability of Piper amalago var.amalago plant as a lime plant substitute for controlling feeding and egglaying behaviors of butterflies.

H. andraemon larvae were reared on Piper amalago var. amalago plant.When they pupated and were ready to lay eggs, the butterflies were giventhe choice of plants on which to lay eggs: unsprayed lime plants,unsprayed Piper amalago var. amalago plants, citrus plant sprayed withPiper amalago var. amalago plant oil, and citrus plant sprayed withd-limonene. The egg laying was permitted to continue until thebutterflies ceased laying eggs. Nearly 100% of the eggs laid hatchedinto larvae.

TABLE 6 Egg laying of Piper amalago var. amalago plant bred H. andraemongiven a choice of plants on which to lay No. of Plants eggs laid % a.Unsprayed Piper amalago var. amalago 133 68.91 plant b. Unsprayed limeplant 8 4.15 c. Citrus plant sprayed with Piper amalago var. 26 13.47amalago plant oil d. Citrus plant sprayed with d-limonene 26 13.47Totals 193 100.00

Table 6 summarizes the results of the experiment in which Piper amalagovar. amalago plant bred butterflies were given the choice of plants onwhich to lay eggs. 68.91% of the eggs were laid on Piper amalago var.amalago plant, establishing that Piper amalago var. amalago plant rearedbutterflies prefer to breed on the type of plant on which they werereared. As expected, few eggs, or 4.15%, were laid on unsprayed limeplant. In total, Piper amalago var. amalago plant reared butterfliesre-laid eggs on Piper amalago var. amalago plants about 96% of the timeand on unsprayed lime plant only about 4% of the time.

This experiment confirmed that butterflies reared on Piper amalago var.amalago plant preferred to lay eggs on Piper amalago var. amalago plantwhen given a choice between sprayed and unsprayed Piper amalago var.amalago plant and both sprayed and unsprayed lime plant. Thus, the useof chemical attractants to induce butterfly egg laying on, e.g., Piperamalago var. amalago plant instead of commercially valuable plants suchas citrus plants, is an effective method for controlling the feeding andbreeding of butterflies.

Example 9

Butterflies were induced to lay on host plant Piper aduncum plantsprayed with lime plant oil, or lime plant sprayed with Piper aduncumplant oil. Piper aduncum shoots were collected in Andrew, Jamaica, and297.35 g of leaves were placed in a Clevenger-type extractor andextracted with boiling water. Once extracted according to the methodsdescribed in Example 2, 0.8919 g of a clear oil with a minty-like odorwas obtained, a yield of 0.30% by weight.

Four Heraclides andraemon pupae were placed in the cage previouslydescribed in Example 1. Three pupae hatched into adults, two males and afemale, which were allowed to mate and lay eggs. They were given achoice of four types of plants on which to lay: unsprayed Piper aduncumcuttings; Piper aduncum cuttings sprayed with d-limonene; unsprayed limeseedlings; and lime seedlings sprayed with the Piper aduncum emulsion.These plants, all with young leaves, were placed around the cage.

Cuttings from the Piper aduncum shoots with young leaves were used toattempt to induce butterflies to feed and lay eggs on a Piper aduncumplant treated with lime plant extract. The Piper aduncum cuttings weresprayed twice daily for eight days, once in the morning and once in theafternoon, with a d-limonene aerosol formulation. The d-limonene aerosolformulation was prepared as a hydrocarbon mixture of 72% propane and 28%butane, with 0.15% d-limonene.

Lime plant seedlings were sprayed with a 5% Piper aduncum emulsion twicedaily for eight days, except no spraying occurred on day five.

As shown in Table 7, the butterflies preferred to lay eggs on unsprayedlime plant seedlings, and did not lay any eggs on unsprayed P. aduncumcuttings or Piper aduncum cuttings sprayed with d-limonene.

After day five, the period during which no Piper aduncum emulsion wassprayed on the lime seedlings, the butterflies, for the first time, tookan interest in the lime seedlings that had previously been sprayed withthe Piper aduncum leaf oil. We observed 9 eggs laid on the young shootsof these lime seedlings that had previously been sprayed with Piperaduncum emulsion. After spraying resumed on day 6, however, thebutterflies again took no interest in the lime seedlings sprayed withPiper aduncum emulsion.

TABLE 7 H. andraemon egg laying on lime plant seedling and P. aduncumNo. of Eggs No. of Eggs Plant after 5 Days after 8 Days % Unsprayed P. 00   0% aduncum cuttings P. aduncum cuttings 0 0   0% sprayed with d-limonene Unsprayed lime 64 79 88.8% seedlings Lime seedlings 9 9 11.1%sprayed with P. aduncum oil. Totals 73 88  100%

Example 10

To determine the toxicity of Piper aduncum leaf essential oil, eightysecond instar H. andraemon larvae were sprayed daily with 2 mL of 2%solution of Piper aduncum leaf essential oils in ethanol with a Potter'stower. For the control, 13 second instar H. andraemon larvae weresprayed daily with 2 mL of ethanol with a Potter's tower.

Seventy-eight of the larvae sprayed with Piper aduncum leaf essentialoils and ethanol died before pupation (97.5% mortality). Two survivinglarvae, both males, pupated after an abnormally lengthy period, 25 days.The two surviving males emerged from pupation after 14 and 16 daysrespectively. Both appeared normal and were able to fly, mate, and lay.

In the control experiment, all 13 larvae pupated after 17 days. Theyappeared normal, except that two had one wing that was slightly smallerthan the other. All 13 adults, however, were able to fly, mate, and lay.

It is to be understood that while the invention has been described inconjunction with the detailed description thereof, the foregoingdescription is intended to illustrate and not limit the scope of theinventions, which are defined by the scope of the appended claims. Otheraspects, advantages, and modifications are within the scope of thefollowing claims.

What is claimed is:
 1. A method of controlling the population size of acitrus-feeding swallowtail butterfly (Heraclides andraemon) specieswhich naturally feeds on a plant of interest, comprising: (a) providinga host plant in a pretreatment state and selecting a chemical attractantextracted from the plant of interest which naturally attract thecitrus-feeding swallowtail butterfly (Heraclides andraemon) species; (b)applying to the host plant the chemical attractant to attract thecitrus-feeding swallowtail butterfly (Heraclides andraemon) species tothe host plant and to cause the citrus-feeding swallowtail butterfly(Heraclides andraemon) species to lay eggs on the host plant and anextract from Piper plant that contains a toxin that kills thecitrus-feeding swallowtail butterfly (Heraclides andraemon) species;wherein a majority of the citrus-feeding swallowtail butterfly(Heraclides andraemon) species or the laid eggs die from ingesting theextract or coming into contact with the extract.
 2. The method of claim1, wherein the chemical attractant is detectable by the swallowtailbutterfly species.
 3. The method of claim 1, wherein application of themixture consists of dipping, spraying, coating, diluting, covering,saturating, misting, fumigating, or dusting the host plant with themixture.
 4. The method of claim 1, wherein the swallowtail butterflyspecies is selected from a group consisting of Heraclides (syn. Papilio)andraemon Hubner, Heraclides andraemon: bonhotei Sharpe, Heraclidesandraemon: andraemon Hubner, Heraclides andraemon: hernandezi de laTorre, Heraclides andraemon: tailori Rothschild, and Heraclidesandraemon: tailori Jordan.
 5. The method of claim 1, wherein the hostplant is a plant on which the swallowtail butterfly species naturallyfeeds in the wild.
 6. The method of claim 1, wherein the host plant is acitrus plant.
 7. The method of claim 1, wherein the host plant is aPiper plant that contains limonene or d-limonene.
 8. The method of claim1, wherein the host plant is one or more plants selected from the groupconsisting of Citrus aurantifolia (Christm.) Swingle (Rutaceae), Piperamalago var. amalago L. (Piperaceae), lime, C. limon (L.) Burm. F.,lemon, C. sinensis (L.) Osbeck, sweet orange, C. reticulata Blanco,Mandarin orange, tangerine, C. paradisi Macf, grapefruit, C. medica L.,citron, C. aurantium L. Seville orange, C. grandis (L.) Osbeck,shaddock, pummelo, C. maxima (Burm.) Merr, ugli, C. reticulata Blanco xC. sinensis (L.) Osbeck, ortanique, C. mitis Blanco, calamondin,Fortunella margarita Lour. Swingle, kumquat (Rutaceae), Amyris P. BrowneRutaceae), Zanthoxylum L., Zanthoxylum martinicense (Lam.), Lantanacamara, Jatropha podagrica, Canna indica, and Z. pterota L. (Rutaceae).9. The method of claim 1, wherein the chemical attractant comprises oilderived from a host plant that contains limonene or d-limonene.
 10. Themethod of claim 1, wherein the chemical attractant comprises oil derivedfrom a citrus plant.
 11. The method of claim 1, wherein the chemicalattractant comprises oil derived from one or more plants selected fromthe group consisting of Citrus aurantifolia (Christm.), Swingle(Rutaceae), Piper amalago var. amalago L. (Piperaceae), lime, C. limon(L.) Burm. F., lemon, C. sinensis (L.) Osbeck, sweet orange, C.reticulata Blanco, Mandarin orange, tangerine, C. paradisi Macf,grapefruit, C. medica L., citron, C. aurantium L. Seville orange, C.grandis (L.) Osbeck, shaddock, pummelo, C. maxima (Burm.) Merr, ugli, C.reticulata Blanco x C. sinensis (L.) Osbeck, ortanique, C. mitis Blanco,calamondin, Fortunella margarita Lour. Swingle, kumquat (Rutaceae),Amyris P. Browne Rutaceae), Zanthoxylum L., Zanthoxylum martinicense(Lam.), Lantana camara, Jatropha podagrica, Canna indica, and Z. pterotaL. (Rutaceae).
 12. The method of claim 1, where in the toxin is derivedfrom Piper aduncum.
 13. The method of claim 1, where in the toxin isapiol.
 14. A method of controlling the population size of acitrus-feeding swallowtail butterfly (Heraclides andraemon) specieswhich naturally feeds on a plant of interest, comprising: (a) providinga host plant in a pretreatment state and selecting a chemical attractantextracted from the plant of interest which naturally attract thecitrus-feeding swallowtail butterfly (Heraclides andraemon) species; (b)applying to the host plant the chemical attractant to attract thecitrus-feeding swallowtail butterfly (Heraclides andraemon) species tothe host plant and to cause the citrus-feeding swallowtail butterfly(Heraclides andraemon) species to lay eggs on the host plant; and (c)applying to the host plant or directly to the citrus-feeding swallowtailbutterfly (Heraclides andraemon) species or the laid eggs an extractfrom Piper plant that contains a toxin that kills the citrus-feedingswallowtail butterfly (Heraclides andraemon) species, wherein a majorityof the citrus-feeding swallowtail butterfly (Heraclides andraemon)species or the laid eggs die from ingesting or coming into contact withthe extract.
 15. The method of claim 14, wherein application of theextract or the chemical attractant consists of dipping, spraying,coating, diluting, covering, saturating, misting, fumigating, or dustingthe host plant with the extract.
 16. The method of claim 14, wherein theswallowtail butterfly species is selected from a group consisting ofHeraclides (syn. Papilio) andraemon Hubner, Heraclides andraemon:bonhotei Sharpe, Heraclides andraemon: andraemon Hubner, Heraclidesandraemon: hernandezi de la Torre, Heraclides andraemon: tailoriRothschild, and Heraclides andraemon: tailori Jordan.
 17. The method ofclaim 14, wherein the host plant is a plant on which the swallowtailbutterfly species naturally feeds in the wild.
 18. The method of claim14, wherein the host plant is a citrus plant.
 19. The method of claim14, wherein the host plant is one or more plants selected from the groupconsisting of Citrus aurantifolia (Christm.) Swingle (Rutaceae), Piperamalago var. amalago L. (Piperaceae), lime, C. limon (L.) Burm. F.,lemon, C. sinensis (L.) Osbeck, sweet orange, C. reticulata Blanco,Mandarin orange, tangerine, C. paradisi Macf, grapefruit, C. medica L.,citron, C. aurantium L. Seville orange, C. grandis (L.) Osbeck,shaddock, pummelo, C. maxima (Burm.) Merr, ugli, C. reticulata Blanco xC. sinensis (L.) Osbeck, ortanique, C. mitis Blanco, calamondin,Fortunella margarita Lour. Swingle, kumquat (Rutaceae), Amyris P. BrowneRutaceae), Zanthoxylum L., Zanthoxylum martinicense (Lam.), Lantanacamara, Jatropha podagrica, Canna indica, and Z. pterota L. (Rutaceae).20. The method of claim 1, where in the toxin is derived from Piperaduncum.
 21. The method of claim 1, where in the toxin is apiol.